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Mnaza Noreen,1 Muhammad Bilal,1,2 Muhammad Usman Qamar,3,4 Nasir Rasool,1 Abid Mahmood,5 Sobia Umar Din,1 Tawaf Ali Shah,6 Yousef A Bin Jardan,7 Mohammed Bourhia,8 Lahcen Ouahmane9 1Department of Chemistry, Government College University Faisalabad, Faisalabad, 38000, Pakistan; 2School of Chemistry and Chemical Engineering, Shandong University, Jinan, 250100, People’s Republic of China; 3Institute of Microbiology, Faculty of Life Sciences, Government College, University Faisalabad, Faisalabad, 38000, Pakistan; 4Division of Infectious Disease and Department of Medicine, University of Geneva, Geneva, Switzerland; 5Department of Pharmaceutical Chemistry, Government College University Faisalabad, Faisalabad, 38000, Pakistan; 6College of Agriculture Engineering and Food Science, Shandong University of Technology, Zibo, 25500, People’s Republic of China; 7Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia; 8Department of Chemistry and Biochemistry, Faculty of Medicine and Pharmacy, Ibn Zohr University, Laayoune, 70000, Morocco; 9Laboratory of Microbial Biotechnologies, Agrosciences and Environment (Biomage), Labeled Research Unit-CNRSTN° 4, Cadi Ayyad University, Marrakesh, 40000, MoroccoCorrespondence: Nasir Rasool; Mohammed Bourhia, Email nasirrasool@gcuf.edu.pk; m.bourhia@uiz.ac.maIntroduction: New Delhi Metallo-β-lactamase producing Klebsiella pneumoniae (NDM-1-KP) sequence type (ST) 147 poses a significant threat in clinical settings due to its evolution into two distinct directions: hypervirulence and carbapenem resistance. Hypervirulence results from a range of virulence factors, while carbapenem resistance stems from complex biological mechanisms. The NDM-1-KP ST147 clone has emerged as a recent addition to the family of successful clones within the species.Methods: In this study, we successfully synthesized 5-bromo-N-alkylthiophene-2-sulfonamides ( 3a-c) by reacting 5-bromothiophene-2-sulfonamide ( 1) with various alkyl bromides ( 2) using LiH. We also synthesized a series of compounds ( 4a-g) from compound ( 3b) using the Suzuki-Miyaura cross-coupling reaction with fair to good yields (56– 72%). Further, we screened the synthesized molecules against clinically isolated New Delhi Metallo-β-lactamase producing Klebsiella pneumoniae ST147. Subsequently, we conducted in-silico tests on compound 3b against a protein extracted from NDM-KP ST147 with PDB ID: 5N5I.Results: The compound ( 3b) with favourable drug candidate status, MIC of 0.39 μg/mL, and MBC of 0.78 μg/mL. This low molecular weight compound exhibited the highest potency against the resistant bacterial strains. The in-silico tests revealed that the compound 3b against a protein extracted from NDM-KP ST147 with PDB ID: 5N5I demonstrated H-bond and hydrophobic interactions.Conclusion: The 5-bromo-N-alkylthiophene-2-sulfonamides displayed antibacterial efficacy against New Delhi Metallo-β-lactamase producing Klebsiella pneumoniae ST147. After the in-vivo trial, this substance might offer an alternative therapeutic option. Keywords: NDM-1, antibacterial, drug resistance, Klebsiella pneumoniae, ST, sulfonamides

Growth patterns and associated endocrine profiles were compared between dominant anovulatory (ADF) and ovulatory follicles (OvF) developing from different waves within and between menstrual cycles in women. Follicular mapping profiles of 49 healthy women of reproductive age and blood samples were obtained every 1–3 days during one interovulatory interval. Sixty-three dominant follicles were classified into wave 1 (W1ADF; n = 8) and wave 2 (W2ADF; n = 6) anovulatory follicles and wave 2 (W2OvF; n = 33) and wave 3 (W3OvF; n = 16) ovulatory follicles. Comparisons were made between W1ADF and W2ADF, W2ADF and W2OvF, and W2OvF and W3OvF. The waves were numbered 1, 2, or 3 based on when the waves emerged relative to the preceding ovulation. W1ADF emerged closer to the preceding ovulation, and W2ADF emerged in the late luteal or early follicular phase. The interval from emergence to maximum diameter was shorter for W2ADF than W1ADF and for W3OvF than W2OvF. Selection of W3OvF occurred at a smaller diameter compared to W2OvF. W1ADF regressed at a faster rate than W2ADF. Also, W1ADF were associated with lower mean follicle stimulating hormone (FSH) and higher mean estradiol than W2ADF. In contrast, W3OvF were associated with higher FSH and luteinizing hormone (LH) compared to W2OvF. However, W2OvF were associated with higher progesterone than W3OvF. This study contributes to the understanding of the physiologic mechanisms underlying selection of the dominant follicle, ovulation, and pathophysiology of anovulation in women, as well as optimization of ovarian stimulation protocols for assisted reproduction.

The current study focused on the exploration of bacterial community in heavy metal contaminated soil using culture dependent and independent approaches that might be applied to the bioremediation process. A total of 150 bacterial colonies were examined from the heavy metal-contaminated soil at initial level, and 25 isolates were chosen for further examination. Finally, twelve strains were chosen for future research based on their high levels of heavy metal resistance. The 10 bacterial strains i.e.,1K-10K were then characterized on the basis of their morphology and microscopic analysis. The morphological and biochemical characteristic relied on the basis of Bergey's manual of bacterial classification that 1K as Pseudomonas sp., 2K as Enterobacter sp., 3K as Streptococcus sp., 4K as Staphylococcus sp.,5K as Staphylococcus sp., 6K as Pseudomonas sp., 7K as Micrococcus sp., 8K as Staphylococcus sp., 9K as Staphylococcus sp. & 10K as Staphylococcus sp. In metagenomics analysis, the most prevalent bacteria present in samples were "Actinobacteria, Proteobacteria, Acidobacteria, Pseudomonas Cyanobacteria, Bacteriodes, and Sphingomonas. Among the 29 identified phyla, Chloroflexi and Tenericutes were the most dominant phyla and their relative abundance ranged from 0.75-1.00. While, on the genera level, the most abundant genera were Blautia, Rhodoplanes followed by Prevotella and Arenimonas. The relative abundance of these genera ranged from 0.75% to 1.00%. Depending on the relative abundance of taxons, the results of this study showed Proteobacteria, Synergistetes and Bacteriodes were most abundant phylum while the least abundant phylum present in heavy metal contaminated soil was Chlamydia and Tenericutes. Screening of soil-based libraries using functional and sequence-based specifications, which has disclosed information on soil microbial communities, has made it possible to identify new microbial communities. Thus, the culture dependent and independent approaches revealed that even heavy metal contaminated soil compose of diverse group of bacterial community that could be explored for bioremediation purposes. [ABSTRACT FROM AUTHOR]

This study emphasized the alpha-amylase gene from Bacillus licheniformis as this species has been witnessed as the most prolific bio-factories for the alpha amylase production on industrial scales with particular reference to its wide commercial applicability. In the present study, a gene encoding alpha-amylase from an indigenously isolated B. licheniformis 208 strain was PCR amplified and sequenced. The obtained 1313 bp long nucleotide sequence (GenBank KM27230) exhibited the closest match with its homologous genes in B. licheniformis ATCC 9945A and ATCC 14580 strains. Gene translation retrieved 437 amino acid long enzyme sequence and exhibits a molecular weight of 50231.8 KDa. Other computed parameters include isoelectric point (5.95), aliphatic index (67.4), instability index (27.45), and GRAVY (-0.627) values. The structure of alphaamylase 208 was found to exhibit a 17 residues long active site, a conserved catalytic site with 3 residues, and a ligand-binding site comprised of 3 residues. Secondary structure configuration in the alpha-amylase 208 was predominated with a-helix and random coils followed by extended strands. The number of ß-turns was found comparatively less. ITASSER assigned E.C number to the BLA 208 sequence was 3.2.1.1 with extracellular origin. Deduced tertiary structure of BLA 208 showed the 7 residues long active site comprising of histidine, arginine, aspartic acid, glutamic acid, tryptophan, histidine, and aspartic acid at position 122, 246, 248, 278, 280, 344, 345 respectively. Na, K, and Ca were observed as the ligand-binding sites existing as a metal triad. Natural selection analysis explains that the alpha-amylase gene in B. licheniformis species is under purifying or negative selection i.e. it stabilizes itself by removing the deleterious mutation. Recombination event analysis was done using GARD (Genetic algorithm for recombination detection). Although, it rather inferred the presence of two break points which were found insignificant when validated via Kishino-Hasegawa test which demonstrates topological incongruence. [ABSTRACT FROM AUTHOR]

The pollution caused by heavy metals is a major problem for our environment, as the high levels of exposure to these metals have harmful consequences for wildlife, vegetation, and human health. Even in trace amounts, several heavy metals, including lead, mercury, cadmium, zinc, arsenic, and nickel, not only possess carcinogenic properties but also have the capacity to cause genetic mutations. In this study, a total of 150 bacteria were isolated, out of which 25 were selected for secondary screening. After secondary screening, five strains were further processed based on their maximum tolerance level. The required indigenous metallotolerant bacterial strains were isolated and identified based on phenotypical and genotypical characterization. The phenotypical characteristics and topology of the phylogram confirm that the bacterial isolate 1LB is Kingella sp., 2LB is Listeria sp., 3LB is Bacillus sp., 4LB is Pseudomonas putida, and 5LB is Cupriavidus necator. According to the result while using LB medium, all the bacterial isolates showed the highest tolerance level against different heavy metal concentrations, i.e., 1LB and 4LB bacterial isolates showed the highest tolerance rate against copper (Cu), while 2LB and 5LB bacterial isolates showed maximum tolerance against chromium (Cr), and 3LB bacterial isolates showed the highest tolerance rate against lead (Pb). Thus, the LB medium was used for the optimization of bioremediation purposes. For bioremediation of heavy metal-contaminated soil, the maximum removal efficiency was 83.80% of the 4LB bacterial strain for Pb, 90.49% of the 5LB bacterial strain for Cu, and 81.87% of the 1LB and 2LB bacterial strains for Cr in the YPG medium. Thus, the results indicate that indigenous heavy metal-tolerant bacterial strains of the area could be used for biological remediation of heavy-metal-contaminated soil, which is the most effective, economical, and environmentally friendly approach as an alternative to traditional methods. [ABSTRACT FROM AUTHOR]

Gastro somatic index and other indices such as Relative Gut Mass (RGM), Relative Length of Gut (RLG), Zihler's Index (ZI) and Gut Vacuity Index (GVI) of Schizothorax labiatus were observed during the present study. The total length of the fish ranged 140mm to 378 mm and Total Weight of fish ranged from 47gm to 449 gm. Lowest GaSI value was observed in the month of April (3.70) and highest value was observed in the month of July (6.81). Annual mean GaSI was recorded as 5.05 ± 0.28. The mean RGM and RLG values observed were 0.03 ± 0.02 and 1.40 ± 0.08 respectively thereby indicating that S. labiatus has omni-herbivore feeding habit. The mean value of Zihler's index (ZI) was recorded as 7.27± 0.24 and mean GVI value was recorded as 9.30 ± 0.65 exhibiting the gluttonous nature of this species. This type of study is helpful in providing important information on food and feeding habits S. labiatus in Kashmir valley. [ABSTRACT FROM AUTHOR]

Citrus fruits, that belong to the Rutaceae. family, have a great economic, nutritional, and therapeutic importance. It has long been a significant source of foreign exchange for Pakistan. Unfortunately, various diseases affect its production and quality, the deadliest of which is known as citrus canker caused by Xanthomonas axonopodis pv. citri. The goal of current research was to isolate and characterize the strains causing citrus canker on the basis of phenotypical and genotypical features. Fruits with possible canker lesions were used for the isolation from the different Citrus Orchards in Khanpur, District Haripur, Pakistan. Citrus fruits, both symptomatic and asymptomatic, were sampled and evaluated during the survey. Only diseased citrus fruits yielded Xanthomonas-like bacterial strains were isolated and identified by carrying out different biochemical tests including catalase, Kovac's oxidase and KOH test. The isolated strains were further identified through polymerase chain reaction (PCR) by using specific ITS primers. Total of eight bacterial isolates were identified to be Xanthomonas spp. through biochemical tests and molecular results revelled the 2A and 2B bacterial isolates as., Xanthomonas citri subsp. citri. (Xcc). Thus, it was concluded that isolated strains were pathogenic bacteria, that is an actual cause of bacterial canker in commercial citrus fruits. Our findings suggested that diseased citrus fruits, could act as a source of dispersal for citrus canker in areas like Pakistan. [ABSTRACT FROM AUTHOR]

OBJECTIVE: The objective of this study was to clone and express the hepatitis B surface antigen gene (HBsAg) in Escherichia coli (E. coli), thereby aiming to develop potential local therapeutics for combating Hepatitis B virus (HBV) infection in the Pakistani community by producing HBsAg in E. coli. MATERIALS AND METHODS: Blood serum samples were collected from hepatitis B-infected patients, and their genomic DNA was extracted. Real-time and nested polymerase chain reaction (PCR) was performed to amplify the HBsAg gene. The gene of interest was cloned into the pET20b expression vector and transformed into E. coli BL21 (DE3) using Isopropyl β-D-1-thiogalactopyranoside (IPTG) induction. The gene’s precise size was confirmed with gene-specific external and internal primers (681 bp and 400 bp, respectively). RESULTS: The HBsAg gene was successfully sequenced and submitted to GenBank, exhibiting 98% homology with targeted HBV sequences worldwide. The expression of HBsAg protein was confirmed through silver staining, Coomassie staining, western blot, and dot blot analysis. CONCLUSIONS: The expressed protein clones are now available for further development as a local recombinant DNA vaccine to prevent hepatitis B viral infection in the local community. [ABSTRACT FROM AUTHOR]

To evaluate the analgesic and physiological effects of electroacupuncture (EA) and medetomindine in domestic goats. Thirty healthy goats (aged 1-2 years and body weight 22±3.17 kg) were divided into five groups (6 goats/group). Group 1 animals were considered as a control; groups 2 and 3 animals were given 5µg kg-1 and 20µg kg-1 medetomidine IM respectively; group 4 animals were stimulated with EA for 30 mins and group 5 animals were given EA in combination with 5µg kg-1 medetomidine IM. The pain threshold in groups 2 and 3 animals were increased significantly (p< 0.05) at 15 to 60 mins compared to group 1. Group 5 animals showed increase (p< 0.05) in the pain threshold at 15 to 60 mins compared to animals in group 2 and group 4. Goats in groups 2 and 3 showed decrease (p< 0.05) in heart rate, respiratory rate, mean arterial pressure and rectal temperature, whereas animals in group 5 showed less effect on these physiological parameters. Furthermore, goats in group 5 did not have any effect on blood cell count, serum chemistry, or liver health status. The combination of EA plus 5 µg kg-1 medetomidine produced adequate antinociception with minimal physiological alteration suggesting that, an effective strategy for clinical observation and minor surgical procedures in domestic goats. [ABSTRACT FROM AUTHOR]

Dengue fever may present with many cardiovascular manifestations contributing to death and disability. Acute Coronary Syndrome (ACS) during dengue is rare but poses important management dilemma. Here we present a series of 5 patients with dengue fever who developed different types of ACS during the ongoing dengue endemic in Bangladesh. Five patients admitted with dengue fever and who developed ACS between 1st July to 30th September 2023 at Medicine Department of Uttara Adhunik Medical College, Dhaka, Bangladesh were studied. All the patients had some comorbidity like diabetes, hypertension, or both. Treatment of ACS was tailored according to their CBC report, most importantly, platelet count, and hematocrit. Four out of the 5 patients responded well to anti-ischemic therapy, even though anti-platelet and anti-coagulants could not be given in all cases. There was 1 fatality, but that was due to ARDS consequent due to severe bilateral pneumonia. Management of ACS in dengue is challenging. Physicians should have appropriate preparedness to deal with these cases. [ABSTRACT FROM AUTHOR]